Division of Science, Research and Environmental Health
Research Project Summary
March 2016
Occurrence of Antibiotic Compounds Found in the Water Column and Bottom
Sediments from a Stream Receiving Two Wastewater Treatment Plant Effluents
in Northern New Jersey
Authors
Jacob Gibs Ph.D. 1
Prepared By
2
R. Lee Lippincott Ph.D. (Project Manager)
Abstract
An urban watershed in northern New Jersey was studied to determine the presence of antibiotic compounds above and
below the discharges of two wastewater treatment plants (WWTP). Many antibiotic compounds are not completely
removed by conventional wastewater treatment plants which results in these compounds entering receiving waters. The
antibiotic compounds which enter waterways from WWTP discharges are assumed to be resistant to biologically mediated
degradation.
Twenty-five antibiotic compounds and six antibiotic compound degradates were analyzed at nine sample locations on two
days in September 2008. Two of the nine sampling locations were background sites upstream of the WWTP discharge
locations on the Hohokus Brook. Another background site was located upstream of a source-water intake for potable water
on the Saddle River above the confluence of the Saddle River and the WWTP discharge receiving stream, the Hohokus
Brook. At low stream flows, the drinking-water source-water intake can receive Hohokus Brook water.
A total of 8 antibiotic compounds and one degradate were detected in the water samples from the sites downstream of the
WWTP discharges. Concentrations of 6 of the 8 detected compounds decreased downstream of the WWTP discharges as
the distance from the WWTP discharges increased. There were no detections of any of the 25 target antibiotic compounds
or six degradates at two of the three background sites in either water or sediment samples. One background site had a
detection of one compound in a sediment sample.
Four compounds were detected in sediments. Two fluoroquinolones and azithromycin were found in their respective
highest concentration at the nearest sampling site downstream of the WWTP discharges. Trimethoprim was only detected
in the sediments of one of the background sites. Pseudo-partition coefficients normalized for sediment organic carbon
concentration were calculated for the three compounds identified in the downstream sediment samples to evaluate the
affinity for certain compounds to partition into the sediment.
Introduction
Two Wastewater Treatment Plants (WWTP) discharge into
the Hohokus Brook, a tributary of the Saddle River. A raw
water intake for drinking water is located on the Saddle
River approximately 1,600 feet upstream of the confluence
of the two streams. Downstream of the confluence is a
weir to maintain a minimum water level for the intake.
Hohokus Brook water can enter the drinking water intake at
stream baseflow levels. The land use of the Saddle River
watershed is 74 percent urban, of which the majority of the
area is sewered (New Jersey Department of Environmental
Protection [NJDEP], 2010). The remaining watershed land
uses are undeveloped and water bodies. Stackelberg et
al. (2007) found that one antibiotic compound and one
antibiotic compound degradate were present in the raw
water of a drinking water treatment plant that was
downstream from several WWTPs. Thus, it is important to
determine the changes in antibiotic compound and degradate
concentrations with distance downstream from WWTP
discharges, and changes in concentrations in stream bottom
sediments which originate from the water column.
The combined Hohokus Brook and Saddle River watershed
was investigated for the presence of antibiotic compounds
because two WWTP discharges are located on Hohokus
Brook and a diversion for potable water is located on the
Saddle River. Under baseflow conditions, the diversion for
potable water has a high probability of collecting the Hohokus
Brook water and the compounds associated with the
subsequent WWTP discharges.
1 Methods
Laboratory Analysis
Sample Collection
Isolation
Samples were collected from nine locations on two days in
September 2008. Three sites upstream of wastewater
treatment plants and drinking water intakes were considered
background for this study. Two of the three background sites
were on the Hohokus Brook and the other was upstream of
the Saddle River-Hohokus Brook confluence. Of the other six
sites, three were on the Hohokus Brook and three were
downstream of the confluence on the Saddle River. Sites 8
and 9 were downstream of the Sprout Brook tributary. Figure
1 shows the study area.
Twenty-five antibiotic compounds and six antibiotic
compound degradates were targeted in this study (Table 1).
Samples were extracted using the HLB Prospekt cartridges
(Waters Corp., Milford, MA). Clinafloxacin, 13C2erythromycin, 13C2-erythromycin-H2O, meclocycline,
simatone, and 13C6-sulfomethoxazole are used as internal
standards while demeclocycline, nalidixic acid,
oleandomycin, and 13C6-sulfamethazine were used as the
surrogate standards. An internal standard solution containing
0.062 ng/µl of simatone and 0.25 ng/µl of each of the other
internal standard was placed in an amber 2-ml
chromatography vial and placed on the LC autosampler tray.
Instrumental Analysis
The laboratory analysis is currently an evolving research
method that was developed at the Kansas Water Science
Center, Organic Geochemistry Research Laboratory that
was modified from a liquid chromatography/tandem mass
spectrometry (LC/MS/MS) version of the on-line solid-phase
extraction (OLSPE) LC/MS method in Meyer and others
(2007). Water samples were analyzed for the analytes listed
in Table 1. LC/MS/MS with electrospray ionization (ESI) used
multiple-reaction monitoring (MRM). Samples were analyzed
in positive-ion mode.
Table 1: Antibiotics Measured In This Research
Antibiotic Class
Field methods for the collection of water-quality data are
described in the USGS National Field Manual (U.S.
Geological Survey, variously dated). A surface-water sample
was collected from the center of flow in an amber, glass bottle
that had been cleaned and baked in the laboratory at 450oF.
Streambed sediment was collected from several areas of
deposition at each site and composited in a methanol-rinsed
stainless steel container. The bed sediments were mixed well
and sieved to retain a less than 2 millimeter fraction. The
stainless steel sieves were cleaned using a soap and water,
de-ionized water rinse, and then methanol-rinsed and air
dried. The sediment samples were placed in a baked, amber
glass jars. All samples were preserved by chilling on ice and
shipped on ice for overnight delivery to the USGS Kansas
Water Science Center, Organic Geochemistry Research
Laboratory.
Reporting Limit (ppb)
Fluoroquinolines
Ciprofloxacin
0.005 µg/L
Enrofloxacin
0.005 µg/L
Lomefloxacin
0.005 µg/L
Norfloxacin
0.005 µg/L
Ofloxacin
0.005 µg/L
Sarafloxacin
0.005 µg/L
Macrolides
Azithromycin
0.005 µg/L
Erythromycin
0.008 µg/L
*Erythromycin-H2O
0.008 µg/L
Roxithromycin
0.005 µg/L
Tylosin
0.010 µg/L
Virginiamycin
0.005 µg/L
Sulfonamides
Sulfachloropyridazine
0.005 µg/L
Sulfadiazine
0.100 µg/L
Sulfadimethoxine
0.005 µg/L
Sulfamethoxazole
0.005 µg/L
Sulfamethazine
0.005 µg/L
Sulfathiazole
0.005 µg/L
Tetracyclines
Chlorotetracycline
0.010 µg/L
Oxytetracycline
0.010 µg/L
Tetracycline
0.010 µg/L
Doxycycline
0.010 µg/L
*Epi-chlorotetracycline
0.010 µg/L
*Epi-iso-chlorotetracycline
0.010 µg/L
*Epi-oxytetracycline
0.010 µg/L
*Epi-tetracycline
0.010 µg/L
*Iso-chlorotetracycline
0.010 µg/L
Miscellaneous antibiotic compounds
Chloramphenicol
0.100 µg/L
Lincomycin
0.005 µg/L
Ormetoprim
0.005 µg/L
Trimethoprim
0.005 µg/L
*Degradation Products
2 Results and Discussions
Conclusion
An abundance of literature reflects the presence of
antibiotics in waterways, especially in waterways receiving
wastewater effluent. The literature also supports the result
that many of the studied antibiotic compounds partition into
the sediments resulting in trace quantities present in the
water column. The results of this investigation were among
the first examples of antibiotic compounds detected in both
the water column and sediment using analytical
methodology developed and refined during this investigation
by the USGS Kansas Water Science Center, Organic
Geochemistry Research Laboratory. Pseudo-partition
coefficients were developed to explain the affinity of certain
antibiotics for the organic carbon compartment in the
sediments. A published journal article describes this
research in more detail (Gibs et al. 2013).
Upstream Background Sites
There were no detections of any of the 25 target antibiotic
compounds or six degradates at the three background sites
in the water column. Trimethoprim was detected in the
sediments at Site 1, a background site. However, this
compound was not detected in the water samples collected
at this site. The detection of trimethoprim currently cannot
be explained using the available data, but its ubiquitous use
as an antibacterial agent could be a possibility for this
random detection. Downstream Water Samples
A total of eight antibiotic compounds (azithromycin,
erythromycin, ciprofloxacin, enrofloxacin, ofloxacin,
trimethoprim, sulfamethazine and sulfamethoazole) and one
degradate (erythromycin-H2O) were detected in the water
samples from the sites downstream of the WWTP
discharges. The concentrations of 6 of the 8 detected
compounds and the one degradate decreased downstream
of the WWTP discharges as the distance from the WWTP
discharges increased. Table 2 summarizes the results of
the water column investigation.
References
Gibs, J., Heckathorn, H., Meyer, M., Klapinski, F., Alebus, M.,
Lippincott, R.L. 2013. Occurrence and partitioning of antibiotic
compounds found in the water column and bottom sediments from
a stream receiving two wastewater treatment plant effluents in
Northern New Jersey, 2008. Science of the Total
Environment.v458-460:107-116.
Lou, Y., Lin, X., Rysz, M., Wang, Y., Zhang, H., and Alvarez, P.J.J.
2011. Occurrence and transport of Tetracycline, Sulfonamide,
Quinolone, and Macrolide Antibiotics in the Haihe River Basin,
China. Environmental Science and Technology, 45:1827-1833.
Downstream Sediments
As previously noted, trimethoprim was detected at one
background site. The other three compounds detected
were azithromycin, a macrolide, and two fluoroquinolines:
ciprofloxacin, and ofloxacin. The highest concentrations of
azithromycin, ciprofloxacin, and ofloxacin detected in
sediments were found at the nearest sampling site located
downstream of the WWTP discharges. The azithromycin
concentration was the highest of the four antibiotics
detected. Table 3 shows the results for the four antibiotic
compounds detected in the stream bottom sediments.
Massey, L.B., Haggard, B.E., Galloway, J.M., Loftin, K.A., Meyer,
M.T., and Green, W.R. 2010. Antibiotic fate and transport in three
effluent-dominated Ozark streams. Ecological Engineering, 36:930938.
Meyer, M.T., Lee, E.A., Ferrell, G.M., Bumgarner, J.E., and Varns,
J. 2007. Evaluation of offline tandem and online solid-phase
extraction with liquid chromatography/electrospray ionization-mass
spectrometry for analysis of antibiotics in ambient water and
comparison to an independent method. U.S. Geological Survey
Scientific Investigations Report 2007-5021, 28 p.
NJ Department of Environmental Protection (NJDEP), Office of
Information Resources Management (OIRM), Bureau of
Geographic Information System (BGIS), 2010. NJDEP 2007 Land
Use/Land Cover Update.
Organic Carbon Concentrations in Sediments
The stream bottom sediment organic carbon concentrations
ranged from 0.6 to 4.4 g/kg (Table 3). Organic-carbon
normalized pseudo-partitioning coefficients (P-PCOC) (Gibs
et al. 2013) were averaged for all sites for an antibiotic
compound when a quantified concentration in water,
sediment, and an organic carbon concentration existed.
The order of the average normalized organic carbon PPCOCs, from highest to lowest was as follows:
(azithromycin) > (ciprofloxacin) ~ (ofloxacin) (Table 4). Lou
et al. (2011) detected 10 antibiotic compounds in the bottom
sediments of the Haihe River, China and estimated the PPC but did not normalize for the organic carbon content. For
comparison, the organic carbon concentrations of the Haihe
River bottom sediments ranged from 15.5 to 38.1 g/kg; an
order of magnitude higher than organic carbon
concentrations found in the sediments of this study. Work
by Massey et al. (2010), which detected nine antibiotic
compounds in the bottom sediments of waterways in
Arkansas, supports the strong relative affinity for the
antibiotics identified in this study to partition into the
sediment.
Stackelberg, P. E., Gibs, J., Furlong, E. T., Meyer, M. T., Zaugg, S.
D., and Lippincott, R. L. 2007. Efficiency of conventional drinkingwater-treatment processes in removal of pharmaceuticals and
other organic compounds. Science of the Total Environment,
377:255-272.
U.S. Geological Survey. variously dated. National Field Manual for
the Collection of Water-Quality Data: U.S. Geological Survey
Techniques of Water-Resources Investigations, book 9, chaps. A1–
A9, available online at http://pubs.water.usgs.gov/twri9A.
This project was funded by the NJDEP, Division of Science,
Research and Environmental Health.
Contract number SR08-050
PRINCIPAL INVESTIGATORS:
1
U.S. Geological Survey PREPARED BY:
2
New Jersey Department of Environmental Protection, Division of
Science, Research and Environmental Health
3 4 RESEARCH PROJECT SUMMARY Please send comments or requests to: Division of Science, Research and Environmental Health Mail code 428‐01, P.O. Box 420 Trenton, NJ 08625 Phone: (609) 984‐6070 Visit the Division of Science, Research & Environmental Health web site @ www.state.nj.us/dep/dsr/ Division of Science, Research & Environmental Health Dr. Gary Buchanan, Director
State of New Jersey Chris ChrisƟe, Governor
5 Department of Environmental Protec on Bob MarƟn, Commissioner